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ABSTRACT The comovirus Bean pod mottle virus (BPMV) is widespread in the soybean-growing regions in the United States. It has a bipartite genome consisting of RNA1 and RNA2, which are encapsidated separately. We previously have reported the occurrence in nature of two distinct subgroups of BPMV strains (subgroups I and II), as well as reassortants between the two subgroups. Here, we report the isolation and molecular characterization of naturally occurring partial diploid reassortant strains, which are diploid for RNA1 and haploid for RNA2. Whereas the RNA1s of the partial diploids are derived from two distinct strain subgroups (I and II), the RNA2 is derived from either subgroup I or II. The partial diploid strains induced strikingly severe symptoms on soybean, which could be explained based on the presence of two distinct RNA1s in the same plant. This conclusion was supported by the finding that pseudo-recombinants constructed with two diverse RNA1s induced very severe symptoms on soybean that mimicked those produced by the naturally occurring partial diploids. No enhancement of symptom severity was observed with pseudorecombinants constructed with closely related RNA1s. Likewise, no enhancement of symptom severity was noted with pseudo-recombinants that are diploid for RNA2 and haploid for RNA1. The potential role of genetic reassortment in the epidemiology and pathogenesis of BPMV is discussed. 相似文献
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Leandro A. Mozzoni Pengyin Chen Rose C. Gergerich 《European journal of plant pathology / European Foundation for Plant Pathology》2009,124(1):133-140
Accessions from Glycine, Phaseolus, and Vigna genera were screened for their reactions to different subgroups of isolates of Bean pod mottle virus (BPMV) in order to establish a differential host system. Screening results indicated that the BPMV isolates differed in pathogenic
aggressiveness but not in virulence. No major resistance genes were found in soybean (Glycine max) or G. soja since all screened accessions showed mosaic or necrotic symptoms to BPMV inoculation. However, these accessions expressed
differences in severity of symptoms when challenged by various BPMV isolates. The inoculation of G. tomentella accessions did not result in mosaic symptoms, and some accessions did not support systemic infection of some of the isolates.
Resistance, presented as a hypersensitive reaction, was observed in some of Phaseolus and Vigna genotypes, and resistant response or susceptibility was stable to all the isolates used in the screening. In conclusion,
the selected G. soja genotypes PI 407019, PI 464889A, and PI 464928, and ‘Amsoy 71’ soybean may help to separate severe (reassortant) from mild
isolates of BPMV based upon their phenotypic reactions. 相似文献
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进口大豆上菜豆荚斑驳病毒的免疫捕获巢式RT-PCR检测 总被引：4，自引：0，他引：4
利用DAS-ELISA对进口大豆上BPMV进行检测发现,从美国进口的部分大豆样品对BPMV的多抗血清呈阳性反应;利用免疫吸附电镜观察发现,ELISA检测阳性的大豆种皮病汁液中存在直径约30nm的球状病毒粒子.根据BPMV外壳蛋白（CP）基因的保守序列设计了2对嵌合引物,建立了BPMV的高灵敏的免疫捕获巢式RT-PCR（IC-nested RT-PCR）检测方法.该方法经免疫捕获、反转录和2轮PCR扩增,能从带毒大豆种子中扩增到预期大小的DNA条带.序列测定与分析表明此条带的序列为BPMV部分CP基因,在系统关系树上与BPMV的其它分离物形成一簇亲缘关系很近.实验表明从进境大豆上检测到了BPMV. 相似文献
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本研究采用DAS ELISA、IC RT PCR及序列测定方法,对加拿大进境大豆种子进行菜豆荚斑驳病毒（Bean pod mottle virus, BPMV）和大豆花叶病毒（Soybean mosaic virus,SMV）检测,结果表明,BPMV的DAS ELISA和IC RT PCR检测结果均为阳性,且PCR扩增产物序列与已报道的BPMV基因序列相似性达97%以上,而SMV的DAS ELISA和IC RT PCR检测结果都为阴性。综合血清学、分子生物学检测结果,确认该批大豆携带有菜豆荚斑驳病毒。 相似文献
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应用TaqMan MGB探针技术检测菜豆荚斑驳病毒 总被引：3，自引：1，他引：2
根据菜豆荚宽驳病毒(Beanpod mottle virus,BPMV)不同分离株外壳蛋白基因(coat protein,CP)的保守序列,设计了特异性引物与TaqMan MGB荧光探针,建立了BPMV的实时荧光RT-PCR检测方法.该方法与酶联免疫检测方法相比,灵敏度提高了100倍,具有快速、灵敏和高特异性的优点,适于对BPMV的快速检测.应用该方法,对来自美国不同地区的携带有BPMV的大豆样品进行检测,均能够得到BPMV的典型扩增曲线,表明引物与探针对BPMV不同分离株具有良好的简并性. 相似文献
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We determined the complete nucleotide sequence of RNA-1 and the 5-terminal region of RNA-2 from Broad bean wilt virus 1 (BBWV-1) isolate PV132. This report is the first analysis of the genome organization of BBWV-1. We also determined the complete nucleotide sequence of RNA-1 from Broad bean wilt virus 2 (BBWV-2) isolate IP and analyzed the genetic relations between BBWV-1 and BBWV-2. Similar to the BBWV-2 isolates, both RNAs of PV132 encoded a single large polyprotein, which was predicted to contain some functional proteins in a manner similar to those of comovirus. With respect to the deduced amino acid sequences of the mature proteins, PV132 and IP had only 20%–40% homology to comovirus. On the other hand, IP was 73%–98% homologous to BBWV-2 isolates, but PV132 was 39%–67% homologous to the isolates. Although the extent of the homologies differed, the homologies were limited between BBWV-1 and BBWV-2 not only for the coat protein but also for the other proteins. These results clearly support the placement of BBWV-1 and BBWV-2 in the genus Fabavirus as distinct species, proposed on the basis of double immunodiffusion tests.The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under the accession numbers AB084450 (RNA-1 of isolate PV132), AB084451 (RNA-2 of isolate PV132), and AB023484 (RNA-1 of isolate IP) 相似文献
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B.C. Lee Y.K. He K. Murao M. Isogai G. Dahal I. Uyeda 《European journal of plant pathology / European Foundation for Plant Pathology》1997,103(6):493-499
Rice dwarf virus isolates were collected from several locations in Japan, the Philippines, China, Nepal and Korea. Genomic dsRNA segment profiles in polyacrylamide gel electrophoresis differed among the isolates. There were less differences in the profiles between isolates from Japan and Korea than in those between these two Countries and others. Nucleic acid hybridization was used to examine the extent of genomic variation. Full-length cDNAs to all genomic segments encoding non-structural proteins (S4, S6, S9, S10, S11 and S12) were synthesized from two Japanese isolates, and were used for dot-blot hybridization. Hybridizations using probes generated from the full-length cDNA clones failed to differentiate isolates from different geographical areas. However, cDNA probes covering a variable region of S12 were able to distinguish Japanese and Korean isolates from those of other countries. Phylogenetic tree analysis based on the amino acid sequence of P12 encoded by S12 grouped Japanese and Korean isolates together. The Chinese isolates from two different locations (Yunnan and Fujian) were closely related to each other, and were the most distantly related to Japanese and Korean isolates. 相似文献
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A varied population of cucumber mosaic virus from peppers 总被引：1，自引：0，他引：1
A varied population of Cucumber Mosaic Virus (CMV) isolates is reported from peppers ( Capsicum annuum ) in Australia. Isolates representing both CMV subgroups (serogroups) I and II have been obtained from the same field at the same sampling time. The CMV isolates were typed into subgroup I (eight isolates) and subgroup II (two isolates) using both a nucleic acid hybridization assay and an immunological assay with monoclonal antibodies. The immunological assay described allows the typing of strains in crude sap extracts, obviating the need for purified virions. The spatial and temporal coincidence of both CMV subgroups presents a situation in which pseudorecombinants with reassorted genomic components might arise. 相似文献
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寻找小菜蛾抗溴氰菊酯品系中差异表达基因,初步分析小菜蛾敏感品系与抗溴氰菊酯品系间的遗传差异.采用改进的cDNA代表性差异分析法,以小菜蛾抗溴氰菊酯品系的cDNA为Driver扩增子、敏感品系的cDNA为Tester扩增子,通过四轮消减杂交,获得1条200bp左右的差异扩增带.将此差异扩增带克隆于pMD18-T载体,随机挑选10个克隆测序.将所得的10个序列与GenBank等数据库作同源比较,发现仅有1个序列与已知泛素基因有较高同源性,但它与小菜蛾抗药性的关系尚未查明. 相似文献
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ABSTRACT Resistance to Soybean mosaic virus (SMV) was identified in PI 88788 soybean, a germ plasm accession from China that is used widely as a source of resistance to soybean cyst nematode. Strains SMV-G1 through -G7 infected the inoculated leaves of PI 88788 but were not detected in upper, noninoculated trifoliolate leaves. Inheritance of resistance was determined by inoculating progenies of crosses of PI 88788 with susceptible cvs. Essex and Lee 68 with SMV strains G1 and G7. Allelomorphic relationships with known genes for resistance to SMV were tested in crosses with the resistant genotypes PI 96983, L29, and V94-5152, possessing Rsv1, Rsv3, and Rsv4 genes, respectively. Data analyses showed that resistance in PI 88788 to SMV-G1 is controlled by a single, partially dominant gene; however, to SMV-G7, the same gene was completely dominant. The PI 88788 gene was independent of the Rsv1 and Rsv3 loci, but allelic to Rsv4 in V94-5152. Expression of the Rsv4 gene in PI 88788 resulted in a reduced number of infection sites and restricted short- and long-distance movement of virus, rather than hypersensitivity. A unique late susceptible phenotype was strongly associated with heterozygosity. This gene has potential value for use in gene pyramiding to achieve resistance to several SMV strains, as well as for rate-reducing resistance. 相似文献
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大豆花叶病毒病研究进展 总被引：15，自引：0，他引：15
大豆花叶病毒病是世界性病害,导致大豆产量降低并产生种粒斑驳。目前国内外对SMV株系的划分不统一。美国报道了G1-G7,G7A,C14九个株系,日本报道了A-E 5个株系,中国东北1、2、3号株系,江苏SA-SH株系,湖北S1,S2株系,黄淮Y 1-Y7株系。各地学者开展了抗源的鉴定和抗病育种工作,筛选和选育出一批抗病品种。美国已命名3个抗性基因,Rsv1,Rsv2,Rsv3。由于抗源不同,对中国东北3个株系SM V抗性遗传研究结果不同,抗性受1对或2对显性或隐性基因控制;对江苏株系抗性遗传研究结果一致,抗性受单显性基因控制。对感染SM V后大豆植株及种粒生理生化性状的变化及抗性机制进行了研究,表明过氧化物酶同工酶等性状与抗性有关。目前已鉴定出与抗性基因连锁的SSR,RFL P和R APD分子标记,成功的克隆了SMV外壳蛋白基因并导入大豆中。 相似文献
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Japanese leaf beet Beta vulgaris var. cicla cv. Fudanso plants were found to contain four double-stranded RNA (dsRNA) components in apparently healthy beet plants. Two were identified as from beet cryptic virus 1 (BCV1), but the other two showed different mobilities on gel electrophoresis and were transcribed into complementary DNA (cDNA) and cloned. Hybridization analysis showed no significant sequence homology between these two dsRNAs and the dsRNA components of BCV1 or the other known cryptic virus of beet, BCV2. Slot- and dot-blot hybridization were used with cDNA clones as probes to identify plants containing these two dsRNA components. Virus particles were purified from these plants and were shown to contain the two new dsRNA components, thus demonstrating the existence of a new beet cryptic virus, which we have called BCV3. 相似文献
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Albiach-Martí MR Guerri J de Mendoza AH Laigret F Ballester-Olmos JF Moreno P 《Phytopathology》2000,90(2):134-138
ABSTRACT A total of 14 Spanish isolates of Citrus tristeza virus (CTV) and 1 isolate from Japan were transmitted by Aphis gossypii, and the subisolates obtained were compared with the source isolates for symptom expression and double-stranded RNA (dsRNA) pattern. Of the 14 Spanish isolates, 9 showed altered dsRNA patterns after aphid transmission but only minor variations in the intensity of symptoms induced on Mexican lime. Northern blot hybridization with complementary DNA (cDNA) probes corresponding to both the 5' and the 3' termini of the CTV genomic RNA (gRNA) showed that the dsRNA bands that could be used to discriminate between the dsRNA pattern of the source and the aphid-transmitted isolates were the replicative forms of defective RNAs (D-RNAs). Conversely, the Japanese isolate and two subisolates obtained from it by aphid transmission had the same dsRNA pattern, but one of the subisolates induced milder symptoms in several hosts. Dot-blot hybridization with cDNA probes representing several regions of the gRNA showed that most of the aphid-transmitted isolates differed from the corresponding source isolate by their hybridization pattern. Our results indicate that aphid transmission often sorts the populations of gRNA variants and D-RNAs present in CTV isolates. 相似文献
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Partial nucleotide sequence analysis of Turkish isolates of Beet necrotic yellow vein virus (BNYVV) RNA-3 总被引：1，自引：0，他引：1
A survey was carried out to detect Beet necrotic yellow vein virus (BNYVV) in soil samples using RT-PCR and bait plant techniques from the sugar beet production area of Tokat, Turkey in 2001. More than 80% of the soil samples analyzed were found to be contaminated with the virus. The partial nucleotide sequence of cDNA corresponding to RNA-3 of BNYVV isolates were analyzed for six different regions of Tokat province. All isolates were assigned to type A strains based on RFLP analysis and DNA sequences. Sequence comparison revealed differences at amino acid positions 35, 68, 71 and 179 of the P25 coding region amongst Turkish isolates. Additionally, all Turkish isolates were compared with Japanese, French, Kazakh, Italian and Belgian isolates. 相似文献
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Genetic Diversity Within Colletotrichum acutatum sensu Simmonds 总被引：1，自引：0，他引：1
ABSTRACT Isolates of Colletotrichum acutatum from several hosts were characterized by various molecular methods in comparison with morphological identification. Species-specific primer analysis was reliable for grouping C. acutatum isolates to their designated species. Arbitrarily primed polymerase chain reaction and A+T-rich DNA analyses identified four subgroups within C. acutatum. Subgroup I contained U.S. isolates from almond, apple, peach, and pecan, subgroup II contained isolates from anemone, olive, and strawberry, subgroup III contained isolates from almond (Israel) and strawberry (Spain), and subgroup IV contained a single isolate from anemone (the Netherlands). Likewise, sequence analysis of the internal transcribed spacer (ITS) 2 region alone or the complete ITS (ITS 1-5.8S-ITS 2) region grouped the isolates into the same four subgroups. Percent similarity of the complete ITS region within each cluster ranged from 99.6 to 100.0, 99.8 to 100.0, and 98.6% among subgroups I, II, and III, respectively. DNA sequence analysis of the ITS 2 region alone or the entire ITS 1-2 region was more informative than that of the ITS 1 region, which could only group the isolates into two main clusters. The molecular methods employed for studying genetic variation in populations of C. acutatum determined that this species is diverse, indicating that isolates within populations of each subgroup are not host specific. 相似文献
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本研究合成了苹果茎沟病毒(ASGV)、苹果褪绿叶斑病毒(ACLSV)和苹果茎痘病毒(ASPV)的生物素标记cDNA探针,对斑点杂交和免疫印迹杂交检测这3种病毒的效果进行了分析。以含ACLSV和ASGV克隆片段的大肠杆菌菌液为样品时,斑点杂交检测灵敏度分别为80 cfu/μL和50 cfu/μL。以离体培养砂梨植株为材料,采用斑点杂交法检测砂梨离体培养植株粗提液中的3种病毒,均产生强的杂交信号,且特异性好。比较斑点杂交和ELISA检测病毒含量相对较低的热处理再生植株中ASGV和ACLSV,结果表明斑点杂交具有较高的灵敏度。组织印迹杂交检测砂梨离体植株ASGV和ACLSV的结果显示,这2种病毒在离体植株的各部位均有分布,自基部至茎尖各部位印迹均产生很强的杂交信号。 相似文献